Different toxic effects of YTX in tumor K-562 and lymphoblastoid cell lines

Yessotoxin (YTX) modulates cellular phosphodiesterases (PDEs). In this regard, opposite effects had been described in the tumor model K-562 cell line and fresh human lymphocytes in terms of cell viability, cyclic adenosine 3´,5´-cyclic monophosphate (cAMP) production and protein expression after YTX treatment. Studies in depth of the pathways activated by YTX in K-562 cell line, have demonstrated the activation of two different cell death types, apoptosis and autophagy after 24 and 48 hours of treatment, respectively. Furthermore, the key role of type 4A PDE (PDE4A) in both pathways activated by YTX was demonstrated. Therefore, taking into account the differences between cellular lines and fresh cells, a study of cell death pathways activated by YTX in a non-tumor cell line with mitotic activity, was performed. The cellular model used was the lymphoblastoid cell line that represents a non-tumor model with normal apoptotic and mitotic machinery. In this context, cell viability and cell proliferation, expression of proteins involved in cell death activated by YTX and mitochondrial mass, were studied after the incubation with the toxin. Opposite to the tumor model, no cell death activation was observed in lymphoblastoid cell line in the presence of YTX. In this sense, variations in apoptosis hallmarks were not detected in the lymphoblastoid cell line after YTX incubation, whereas this type I of programmed cell death was observed in K-526 cells. On the other hand, autophagy cell death was triggered in this cellular line, while other autophagic process is suggested in lymphoblastoid cells. These YTX effects are related to PDE4A in both cellular lines. In addition, while cell death is triggered in K-526 cells after YTX treatment, in lymphoblastoid cells the toxin stops cellular proliferation. These results point to YTX as a specific toxic compound of tumor cells, since in the non-tumor lymphoblastoid cell line, no cell death hallmarks are observed

Transcript expression of vesicular glutamate transporters in lumbar dorsal root ganglia and the spinal cord of mice – Effects of peripheral axotomy or hindpaw inflammation

Using specific riboprobes, we characterized the expression of vesicular glutamate transporter (VGLUT)1–VGLUT3 transcripts in lumbar 4–5 (L4–5) dorsal root ganglions (DRGs) and the thoracolumbar to lumbosacral spinal cord in male BALB/c mice after a 1- or 3-day hindpaw inflammation, or a 7-day sciatic nerve axotomy. Sham animals were also included. In sham and contralateral L4–5 DRGs of injured mice, VGLUT1-, VGLUT2- and VGLUT3 mRNAs were expressed in ∼45%, ∼69% or ∼17% of neuron profiles (NPs), respectively. VGLUT1 was expressed in large and medium-sized NPs, VGLUT2 in NPs of all sizes, and VGLUT3 in small and medium-sized NPs. In the spinal cord, VGLUT1 was restricted to a number of NPs at thoracolumbar and lumbar segments, in what appears to be the dorsal nucleus of Clarke, and in mid laminae III–IV. In contrast, VGLUT2 was present in numerous NPs at all analyzed spinal segments, except the lateral aspects of the ventral horns, especially at the lumbar enlargement, where it was virtually absent. VGLUT3 was detected in a discrete number of NPs in laminae III–IV of the dorsal horn. Axotomy resulted in a moderate decrease in the number of DRG NPs expressing VGLUT3, whereas VGLUT1 and VGLUT2 were unaffected. Likewise, the percentage of NPs expressing VGLUT transcripts remained unaltered after hindpaw inflammation, both in DRGs and the spinal cord. Altogether, these results confirm previous descriptions on VGLUTs expression in adult mice DRGs, with the exception of VGLUT1, whose protein expression was detected in a lower percentage of mouse DRG NPs. A detailed account on the location of neurons expressing VGLUTs transcripts in the adult mouse spinal cord is also presented. Finally, the lack of change in the number of neurons expressing VGLUT1 and VGLUT2 transcripts after axotomy, as compared to data on protein expression, suggests translational rather than transcriptional regulation of VGLUTs after injury.Fil: Malet, Mariana. Universidad Austral. Facultad de Ciencias Biomédicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Vieytes, C. A.. Universidad Austral. Facultad de Ciencias Biomédicas; ArgentinaFil: Lundgren, K. H.. University of Cincinnati; Estados UnidosFil: Seal, R. P.. University of Pittsburgh; Estados UnidosFil: Tomasella, María Eugenia. Universidad Austral. Facultad de Ciencias Biomédicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Seroogy, K. B.. University of Cincinnati; Estados UnidosFil: Hökfelt, T.. Karolinska Huddinge Hospital. Karolinska Institutet; SueciaFil: Gebhart, G. F.. University of Pittsburgh; Estados UnidosFil: Brumovsky, Pablo Rodolfo. Universidad Austral. Facultad de Ciencias Biomédicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. University of Pittsburgh; Estados Unido

Furanoditerpenes from Spongia (Spongia) tubulifera Display Mitochondrial-Mediated Neuroprotective Effects by Targeting Cyclophilin D

[Abstract] Neuroprotective properties of five previously described furanoditerpenes 1–5, isolated from Spongia (Spongia) tubulifera, were evaluated in an in vitro oxidative stress model in SH-SY5Y cells. Dose–response treatments revealed that 1–5 improved cell survival at nanomolar concentrations through the restoration of mitochondrial membrane potential and the reduction of reactive oxygen species. Their ability to prevent the mitochondrial permeability transition pore opening was also assessed, finding that 4 and 5 inhibited the channel at 0.001 μM. This inhibition was accompanied by a decrease in the expression of cyclophilin D, the main regulator of the pore, which was also reduced by 1 and 2. However, the activation of ERK and GSK3β, upstream modulators of the channel, was not affected by compounds. Therefore, their ability to bind cyclophilin D was evaluated by surface plasmon resonance, observing that 2–5 presented equilibrium dissociation constants in the micromolar range. All compounds also showed affinity for cyclophilin A, being 1 selective toward this isoform, while 2 and 5 exhibited selectivity for cyclophilin D. When the effects on the intracellular expression of cyclophilins A–C were determined, it was found that only 1 decreased cyclophilin A, while cyclophilins B and C were diminished by most compounds, displaying enhanced effects under oxidative stress conditions. Results indicate that furanoditerpenes 1–5 have mitochondrial-mediated neuroprotective properties through direct interaction with cyclophilin D. Due to the important role of this protein in oxidative stress and inflammation, compounds are promising drugs for new therapeutic strategies against neurodegeneration.The research leading to these results has received funding from the following FEDER cofunded grants: from Conselleria de Cultura, Educacion e Ordenación Universitaria, Xunta de Galicia, GRC (ED431C 2021/01), and GRC2018/039; from the Ministerio de Ciencia e Innovación IISCIII/PI19/001248 and PID 2020-11262RB-C21; from European Union Interreg AlertoxNet EAPA-317-2016 and Interreg Agritox EAPA-998-2018 and H2020 778069-EMERTOX; and from BLUEBIOLAB (0474_BLUEBIOLAB_1_E), Programme INTERREG V A of Spain-Portugal (POCTEP). R.A. was supported by a postdoctoral fellowship from Xunta de Galicia (ED481B-2021-038), Spain. D.P-P. received a postdoctoral fellowship from the National Council of Science and Technology (CONACYT) of MexicoXunta de Galicia; ED431C 2021/01Xunta de Galicia; GRC2018/039Xunta de Galicia; ED481B-2021-03

Validación experimental de un modelo que describe la etapa axial de un plasma focus sin electrodo externo

Se presenta una modificación al modelo de la barredora de nieve tradicional donde se supone que la lámina se forma sobre el aislante y tiene un espesor , pudiendo luego evolucionar tanto en la dirección radial como en la axial. La evolución se realiza planteando la ecuación de conservación de la cantidad de movimiento en la cual, para tener en cuenta la curvatura de la lámina, se agrega un segundo parámetro (eficiencia de transferencia de impulso, ) en el término que contabiliza la cantidad de movimiento cedida al gas barrido. El modelo incluye la ecuación del circuito eléctrico y el sistema de ecuaciones resultante se resuelve numéricamente. Los parámetros libres se ajustan a partir de mediciones de la cinemática de la lámina obtenidas con una sonda magnética inductiva en un equipo Plasma Focus con un electrodo central de 9,5 mm de diámetro y 90 mm de longitud. El segundo electrodo está formado por un disco plano sobre el que asienta el aislante y el electrodo central. Los gases empleados fueron hidrógeno y nitrógeno, en el rango 0,5 – 10 mbar. Se encuentra que tanto como decrecen al aumentar la presión del gas de llenado.An extension to the standard Snow Plow model is presented which assumes that the current sheath is formed along the insulator with a width , and it is then able to evolve both in the axial and radial directions. This evolution is modelled taking into account the conservation of momentum, but a second parameter is included (momentum transfer efficiency ) in order to consider the curvature of the current sheath. The model also includes an equation for the electric circuit. The resulting set of equations is solved numerically. Both parameters are fitted using measurements of sheath cinematics taken with magnetic probes. A Plasma Focus device with a central electrode of 9.5 mm in diameter, 90 mm long and with no outer electrode was used. The filling gas was either hydrogen or nitrogen and the filling pressure was varied in the range 0.5 – 10 mbar. We found that both and decrease as the filling gas pressure is increased.ISSN 0327-358

Improvements in the determination of ISS Ca II K parameters

Measurements of the ionized Ca II K line are one of the major resources for long-term studies of solar and stellar activity. They also play a critical role in many studies related to solar irradiance variability, particularly as a ground-based proxy to model the solar ultraviolet flux variation that may influence the Earth's climate. Full disk images of the Sun in Ca II K have been available from various observatories for more than 100 years and latter synoptic Sun-as-a-star observations in Ca II K began in the early 1970s. One of these instruments, the Integrated Sunlight Spectrometer (ISS) has been in operation at Kitt Peak (Arizona) since late 2006. The ISS takes daily observations of solar spectra in nine spectra bands, including the Ca II K and H line s. We describe recent improvements in data reduction of Ca II K observations, and present time variations of nine parameters derived from the profile of this spectral line

Pulsar wind nebulae around the southern pulsars PSR B1643-43 and PSR B1706-44

We present high resolution VLA images taken at the wavelengths of lambda 20 cm, lambda 6 cm, and lambda 3.6 cm in the vicinity of the pulsars PSR B1706-44 (PSR J1709-4428) and PSR B1643-43 (PSR J1646-4346). Both of these pulsars are young (<30,000 yrs) and have large spin-down luminosities (e+35 erg/sec) and hence are good candidates to search for extended synchrotron nebula excited by the relativistic pulsar wind. For PSR B1643-43 we found evidences of a 4'comet-shaped nebula, suggestive of a synchrotron ``wake'' left by a fast moving pulsar. PSR B1706-44 appears surrounded by a spherical nebula approximately 3 ' in diameter. Based on their morphology, the detection of significant linear polarization (>20%), and their flat radio spectra (alpha=0.25-0.3, where S_nu aprox nu^-alpha) we argue that these are wind nebulae powered by the rotational energy loss of the respective pulsars

Crambescin C1 Acts as A Possible Substrate of iNOS and eNOS Increasing Nitric Oxide Production and Inducing In Vivo Hypotensive Effect

Crambescins are guanidine alkaloids from the sponge Crambe crambe. Crambescin C1 (CC) induces metallothionein genes and nitric oxide (NO) is one of the triggers. We studied and compared the in vitro, in vivo, and in silico effects of some crambescine A and C analogs. HepG2 gene expression was analyzed using microarrays. Vasodilation was studied in rat aortic rings. In vivo hypotensive effect was directly measured in anesthetized rats. The targets of crambescines were studied in silico. CC and homo-crambescine C1 (HCC), but not crambescine A1 (CA), induced metallothioneins transcripts. CC increased NO production in HepG2 cells. In isolated rat aortic rings, CC and HCC induced an endothelium-dependent relaxation related to eNOS activation and an endothelium-independent relaxation related to iNOS activation, hence both compounds increase NO and reduce vascular tone. In silico analysis also points to eNOS and iNOS as targets of Crambescin C1 and source of NO increment. CC effect is mediated through crambescin binding to the active site of eNOS and iNOS. CC docking studies in iNOS and eNOS active site revealed hydrogen bonding of the hydroxylated chain with residues Glu377 and Glu361, involved in the substrate recognition, and explains its higher binding affinity than CA. The later interaction and the extra polar contacts with its pyrimidine moiety, absent in the endogenous substrate, explain its role as exogenous substrate of NOSs and NO production. Our results suggest that CC serve as a basis to develop new useful drugs when bioavailability of NO is perturbed.Fil: Rubiolo, Juan Andrés. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina. Ministerio de Ciencia, Tecnologia E Innovacion Productiva (santa Fe). - Gobierno de la Provincia de Santa Fe. Ministerio de Ciencia, Tecnologia E Innovacion Productiva (santa Fe).; Argentina. Universidad de Santiago de Compostela; España. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Lence, Emilio. Universidad de Santiago de Compostela; EspañaFil: González Bello, Concepción. Universidad de Santiago de Compostela; EspañaFil: Roel, María. Universidad de Santiago de Compostela; EspañaFil: Gil Longo, José. Universidad de Santiago de Compostela; EspañaFil: Campos Toimil, Manuel. Universidad de Santiago de Compostela; EspañaFil: Ternon, Eva. Université Nice Sophia Antipolis. Laboratoire Jean-alexandre Dieudonné.; FranciaFil: Thomas, Olivier P.. National University of Ireland Galway; IrlandaFil: González Cantalapiedra, Antonio. Universidad de Santiago de Compostela; EspañaFil: López Alonso, Henar. Universidad de Santiago de Compostela; EspañaFil: Vieytes, Mercedes R.. Universidad de Santiago de Compostela; EspañaFil: Botana, Luis M.. Universidad de Santiago de Compostela; Españ

Psammaplin A and Its Analogs Attenuate Oxidative Stress in Neuronal Cells through Peroxisome Proliferator-Activated Receptor γ Activation

Psammaplins are sulfur containing bromotyrosine alkaloids that have shown antitumor activity through the inhibition of class I histone deacetylases (HDACs). The cytotoxic properties of psammaplin A (1), the parent compound, are related to peroxisome proliferator-activated receptor γ (PPARγ) activation, but the mechanism of action of its analogs psammaplin K (2) and bisaprasin (3) has not been elucidated. In this study, the protective effects against oxidative stress of compounds 1–3, isolated from the sponge Aplysinella rhax, were evaluated in SH-SY5Y cells. The compounds improved cell survival, recovered glutathione (GSH) content, and reduced reactive oxygen species (ROS) release at nanomolar concentrations. Psammaplins restored mitochondrial membrane potential by blocking mitochondrial permeability transition pore opening and reducing cyclophilin D expression. This effect was mediated by the capacity of 1–3 to activate PPARγ, enhancing gene expression of the antioxidant enzymes catalase, nuclear factor E2-related factor 2 (Nrf2), and glutathione peroxidase. Finally, HDAC3 activity was reduced by 1–3 under oxidative stress conditions. This work is the first description of the neuroprotective activity of 1 at low concentrations and the mechanism of action of 2 and 3. Moreover, it links for the first time the previously described effects of 1 in HDAC3 and PPARγ signaling, opening a new research field for the therapeutic potential of this compound family

The relative emission from chromospheres and coronae: Dependence on spectral type and age

Extreme-ultraviolet and X-ray emission from stellar coronae drives mass loss from exoplanet atmospheres, and ultraviolet emission from stellar chromospheres drives photochemistry in exoplanet atmospheres. Comparisons of the spectral energy distributions of host stars are, therefore, essential for understanding the evolution and habitability of exoplanets. The large number of stars observed with the MUSCLES, Mega-MUSCLES, and other recent Hubble Space Telescope observing programs has provided for the first time a large sample (79 stars) of reconstructed Lyα fluxes that we compare with X-ray fluxes to identify significant patterns in the relative emission from these two atmospheric regions as a function of stellar age and effective temperature. We find that as stars age on the main sequence, the emissions from their chromospheres and coronae follow a pattern in response to the amount of magnetic heating in these atmospheric layers. A single trend-line slope describes the pattern of X-ray versus Lyα emission for G and K dwarfs, but the different trend lines for M dwarf stars show that the Lyα fluxes of M stars are significantly smaller than those of warmer stars with the same X-ray flux. The X-ray and Lyα luminosities divided by the stellar bolometric luminosities show different patterns depending on stellar age. The L (Lyα)/L(bol) ratios increase smoothly to cooler stars of all ages, but the L(X)/L(bol) ratios show different trends. For older stars, the increase in coronal emission with decreasing Teff is much steeper than that of chromospheric emission. We suggest a fundamental link between atmospheric properties and trend lines relating coronal and chromospheric heating.Fil: Linsky, Jeffrey L.. State University of Colorado at Boulder; Estados UnidosFil: Wood, Brian E.. Spece Sciences División. Naval Research Laboratory; Estados UnidosFil: Youngblood, Allison. State University of Colorado at Boulder; Estados UnidosFil: Brown, Alexander. State University of Colorado at Boulder; Estados UnidosFil: Froning, Cynthia S.. University of Texas at Austin; Estados UnidosFil: France, Kevin. State University of Colorado at Boulder; Estados UnidosFil: Buccino, Andrea Paola. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Astronomía y Física del Espacio. - Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Astronomía y Física del Espacio; ArgentinaFil: Cranmer, Steven R.. State University of Colorado at Boulder; Estados UnidosFil: Mauas, Pablo Jacobo David. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Astronomía y Física del Espacio. - Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Astronomía y Física del Espacio; ArgentinaFil: Miguel, Yamila. Leiden Observatory; Países BajosFil: Sebastian Pineda, J.. State University of Colorado at Boulder; Estados UnidosFil: Rugheimer, Sarah. University of Oxford; Reino UnidoFil: Vieytes, Mariela Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Astronomía y Física del Espacio. - Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Astronomía y Física del Espacio; ArgentinaFil: Wheatley, Peter J.. University of Warwick; Reino UnidoFil: Wilson, David J.. University of Texas at Austin; Estados Unido

Toxic Action Reevaluation of Okadaic Acid, Dinophysistoxin-1 and Dinophysistoxin-2: Toxicity Equivalency Factors Based on the Oral Toxicity Study

Background/Aims: Okadaic acid (OA) and the structurally related compounds dinophysistoxin-1 (DTX1) and dinophysistoxin-2 (DTX2) are marine phycotoxins that cause diarrheic shellfish poisoning (DSP) in humans due to ingestion of contaminated shellfish. In order to guarantee consumer protection, the regulatory authorities have defined the maximum level of DSP toxins as 160 µg OA equivalent kg-1 shellfish meat. For risk assessment and overall toxicity determination, knowledge of the relative toxicities of each analogue is required. In absence of enough information from human intoxications, oral toxicity in mice is the most reliable data for establishing Toxicity Equivalence Factors (TEFs). Methods: Toxins were administered to mice by gavage, after that the symptomatology and mice mortality was registered over a period of 24 h. Organ damage data were collected at necropsy and transmission electron microscopy (TEM) was used for ultrastructural studies. Toxins in urine, feces and blood were analyzed by HPLC-MS/MS. The evaluation of in vitro potencies of OA, DTX1 and DTX2 was performed by the protein phosphatase 2A (PP2A) inhibition assay. Results: Mice that received DSP toxins by gavage showed diarrhea as the main symptom. Those toxins caused similar gastrointestinal alterations as well as intestine ultrastructural changes. However, DSP toxins did not modify tight junctions to trigger diarrhea. They had different toxicokinetics and toxic potency. The lethal dose 50 (LD50) was 487 µg kg-1 bw for DTX1, 760 µg kg-1 bw for OA and 2262 µg kg-1 bw for DTX2. Therefore, the oral TEF values are: OA = 1, DTX1 = 1.5 and DTX2 = 0.3. Conclusion: This is the first comparative study of DSP toxins performed with accurate well-characterized standards and based on acute toxicity data. Results confirmed that DTX1 is more toxic than OA by oral route while DTX2 is less toxic. Hence, the current TEFs based on intraperitoneal toxicity should be modified. Also, the generally accepted toxic mode of action of this group of toxins needs to be reevaluated